Means and method for harvesting and handling tissue samples for biopsy analysis

ABSTRACT

A tissue biopsy sample is placed on a tissue trapping and supporting material that can withstand tissue preparation procedures and which can be cut with a microtome. The tissue is immobilized on the material and the material and the tissue are subjected to a process for replacing tissue fluids with wax, and then the tissue and the supporting material are sliced for mounting on slides using a microtome. Harvesting devices and containers using the filter material are disclosed. An automated process is also disclosed. One form of the invention has the tissue trapping and supporting material porous and another form of the invention includes a tissue supporting material that is not easily microtomed.

TECHNICAL FIELD OF THE INVENTION

The present invention relates to the general art of analysis of tissuesamples, and to the particular field of obtaining, handling andprocessing tissue biopsy samples.

BACKGROUND OF THE INVENTION

When disease is suspected in a living being, the physician must arriveat a specific diagnosis. Some disease processes, particularly tumors,require a histologic and/or cytologic diagnosis. While radiologic toolsare useful in detecting the presence of a tumor, the cell type of thetumor can only be determined by a pathologist's examination of ahistologic or cytologic sample of the tumor. There are a number ofdevices that have been fashioned to actually perform the act of takingtissue samples. These devices may obtain tissue for histology or in thecase of needle aspiration biopsies, samples for cytology and histology.In many cases, these samples are very small and difficult to retrieveand process. These small tissue fragments may originate from a punch, orsimilar biopsy procedure devices or from Fine Needle Aspiration Biopsy(FNAB) biopsies. FNAB is typical and produces single cells, small cellclumps and fragments which are immediately smeared onto a glass slide(direct smears) or rinsed into a container with preservative fluid.After being transported to the laboratory, these samples are centrifugedonto a glass slide (cytospin smears). In some cases needle aspirationbiopsy produces tissue fragments which are large enough to processhistologically. If successfully retrieved, these fragments are submittedin blood clot or agar in a technique known as cell block preparationwhich are then immobilized in wax for sectioning and slide preparation.

FNAB is one example of the tissue collection techniques used and theproblems which are of interest to the present invention. Fine NeedleAspiration Biopsy techniques have been practiced for many years and theliterature contains many studies on technique and comparison of variousimproved devices for same. There exists two different kinds of biopsyneedles. Those with active or movable cutting elements and those thatare passive or non-moving. Active needles have two basic problems, whichare cost and complexity. The needles that are of interest to thisinvention are most often 22 gauge which is 0.028" OD with a standardwall of 0.006". This leaves only 0.016" ID. Some prior art designs usean active element down the ID bore to sever and capture tissue. 0.016"does not provide a great deal of clearance for these elements and thusthese prior art needles are inefficient. If it is desired to furthersuck tissue fragments up the needle bore further reducing the bore, thebore will be further reduced because a second element must be added,which is counter productive.

Other methods of obtaining samples are also discussed in the literatureand also have problems. Each is characterized by tissue size and numberof pieces generally available as well as whether orientation in theeventual sectioning plane is critical for example:

Fine Needle Aspiration Biopsy--very small pieces of tissue taken fromthe core of a fine needle; usually transported in fixative solution;

GI biopsy--characterized by a few small tissue pieces; it is desirableto concentrate the tissue pieces in close proximity to each other;

Prostate chips--orientation is irrelevant for these samples;

Endometrial Curettings--characterized by varying size samples;orientation is irrelevant;

Vessel--orientation is critical; sections need to be transverse;

Core Biopsy--i.e. from the prostate--orientation is critical; the tissueshould lie flat all in the same plane;

Gall bladder--orientation is critical--the tissue should be embedded onedge;

Uterine Wall, breast or large tumors--orientation is notcritical--sample lies flat in a plane.

Some of these methods are characterized by the possibility of supplyingextremely small tissue samples. Some samples can be as small as a fewcells, and extremely small samples can create problems. These problemsinclude loss of the sample, dehydration of the sample, and contaminationof the sample during harvesting, storage and transport. Still further,as will be more evident from the following discussion, small samples areextremely difficult and time consuming to process in the laboratory.

Still further, in many cases, a tissue sample is mixed with effluent.Prior art devices and methods account for collection of effluent onlyand do not provide means and methods for trapping tissue specimens. Theprior art collects effluent, but does not provide means or methods forthe separation of tissue from the effluent. Therefore, there is a needfor a means and a method for handling effluent as well as tissue samplesand for efficiently separating tissue from effluent.

Once a tissue sample is harvested it must be transported to thepathology lab for processing. Currently, handling and processing ofsmall biopsies in the histology laboratory is a tedious task andrequires multiple manual manipulations of the specimen. Fine NeedleAspiration Biopsy (FNAB) is typical. Therefore, there is a need tohandle and process very small samples of tissue in an expeditiousmanner.

In addition to the above problems, a further problem with currently usedmeans and methods is associated with the orientation of samples.Currently, in a pathology lab, the pathologist will gross-in the tissuesamples, cut them into appropriate size specimens, if necessary, andplace them into a tissue cassette for processing. Herein lies one of thebiggest problems of the existing art. When the tissue sample is placedinto the tissue cassette, the pathologist orients the sample so that anysurface in which he or she desires to see sectioned is placed face up inthe cassette. The histotech who retrieves the tissue from the cassetteafter processing knows through training that when opening the cassettethe tissue surface that faces up when first opened is then placed facedown into the wax mold, which in turn will become the first surface tobe sectioned by a microtome blade. This is an established protocol whichis observed in most pathology labs today. This process then necessitateshuman involvement and redundant handling. In addition, sometimes specialsponge materials must be packed into the cassette to keep a sampleoriented or to prevent loss from the cassette if it is too small and mayturn or lose its orientation during the tissue processing. Sometimes,notes and drawings accompany tissue samples to show how they should beoriented in the wax.

No current system or method provides the ability to maintain criticaltissue orientation throughout these steps and eliminate human errors inthe associated manual steps and procedures. Therefore, there is a needfor a system and a process that can maintain the preferred orientationof the tissue sample from the time of initial gross-in throughout thetissue processing procedure and continuing through the wax embeddingstage with no human involvement required beyond initial gross in.

Another problem encountered with presently available systems is the lackof integration and multiple handling steps required to produce asectioned sample for pathological examination. Therefore, there is aneed for an approach which reduces the time and handling of biopsysamples.

By way of background, a review of the standard procedure that eachsample must undergo to get from harvest to a prepared histologic slideis necessary. First, the sample must be taken with the appropriateinstrument. The tissue is then retrieved from the instrument anddeposited into some sort of specimen container, usually with a fixativesuch as 10% formalin. The container is labeled and transported to thepathology lab. Herein lies the first problem with the prior art. With noway to control where the sample lodges in the container, the sample maystick to the lid or sides of the container and become dried out beforeit reaches the pathology lab; rendering it difficult, if not impossibleto interpret. In addition, the samples may be extremely small and may behard to locate and retrieve from the container.

When the pathology laboratory receives the container, the specimen islogged into the manual or computerized anatomic pathology system and isassigned a unique surgical pathology accession number. This number isplaced on the specimen container and is subsequently used to labelhistology slides, cassettes and the final surgical pathology report. Thespecimen is logged into the paperwork system and physically described inan appropriate medium, such as dictation or the like, by a pathologistor assistant. This is the description portion of the process known as"grossing-in" the specimen. The grossing in continues when thepathologist or assistant manually retrieves the specimen and views thespecimen, and then sections the specimen into appropriate size morsels,if necessary, and places them into a plastic tissue cassette. If verytiny or multiple, the pieces of tissue must be immobilized within somedevice such as two layers of sponge or a tea bag to prevent them fromescaping from the cassette during processing. Many times a surgeon willhave taken diffuse biopsy samples or scrapings from the mucosal liningof an organ, such as an endocervical biopsy. Often these samples arevery small and multiple such as is the case with tissue fragments fromFine Needle Aspiration Biopsy (FNAB). Other times a doctor will depositthe sample in filter paper which resembles a tea bag. All of thesevarious tissue specimens end up in a tissue cassette. As used herein,the term "grossing-in" includes both the description of the tissuesample and the preparation of the tissue sample for further processing.

At the end of the day all of the cassettes are put into a tissueprocessor where the tissue is subjected to a sequence of solutions andheat. These solutions gradually replace water in the cells with alcohol,followed by xylene, and ultimately by wax. This gives thewax-impregnated tissue a similar consistency to the wax surrounding thetissue in the next step. After the tissue processing is complete,usually the following morning, the sample is again handled to remove itfrom the cassette where it is placed and oriented in a mold. At thispoint if a tea bag or sponge was used to immobilize the sample, thepathology lab is then faced with trying to extract or scrape thewax-impregnated specimen from the paper, before placing the specimen inthe wax mold.

Hot (molten) paraffin wax is poured into the mold to immobilize thetissue in a solid block of wax. After cooling, the wax block is removedfrom the mold, placed into a microtome and sectioned into thin slicesapproximately 4--6 microns thick. These sections are floated onto glassslides, stained, cover-slipped, and are then ready for microscopicexamination. In this process, samples are handled or transferred manytimes. Each handling process takes time and human involvement.

Therefore, there is a need for a means and method to improve theharvesting of tissue samples. There is also a need for handling andprocessing those harvested tissue samples in an efficient and reliablemanner that lends itself to automation and removes the need for a humanto find, handle and orient a tissue sample before analysis of thatsample can be performed.

OBJECT OF THE INVENTION

It is a main object of the present invention to provide a means andmethod for handling harvested tissue samples in an efficient mannerwhich lends itself to automation.

It is another object of the present invention to provide a system and aprocess that can maintain the preferred orientation of the tissue samplefrom the time of initial gross-in throughout the tissue processingprocedure and continuing through the wax embedding stage with no humaninvolvement required beyond initial gross-in.

It is another object of the present invention to provide a means and amethod for efficiently harvesting tissue samples for biopsy.

It is another object of the present invention to provide a means andmethod for handling harvested tissue samples in an efficient manner witha minimum of human intervention.

It is another object of the present invention to provide tissue trappingand supporting means that can retain tissue samples and facilitate easytransfer of the specimen without having to individually retrieve smalltissue fragments from a sample container.

It is another object of the present invention to provide a tissuetrapping sectionable supporting means or stage that is constructed of amaterial that is able to be sectioned in a microtome and appearsnon-distracting in the histologic sections and does not stain withtissue stains applied to the sections.

It is another object of the present invention to provide a tissuetrapping platform that is constructed of a material that is imperviousto the harsh chemical and temperature environment of a tissue waxprocessor machine.

It is another object of the present invention to provide a tissuetrapping platform that is constructed of a material that is imperviousto the chemical and temperature environment of a tissue wax embeddingmachine and may have a surface modification improving wettability on thefilter or stage of the platform. The stages may be sectionable or not.

It is another object of the present invention to provide a biopsycontainer that holds the specimen trapping sectionable means for easyplacement of tissue samples, and assures that the tissue remainscontinually submerged in the fixative solution and further allows theremoval of the tissue trapping and supporting means and specimen withease.

It is another object of the present invention to provide a method forimmobilizing the tissue on a trapping platform to facilitate automationof the wax embedding process.

It is another object of the present invention to provide a method ofautomating the cell block tissue preparation, processing and waxembedding procedures.

It is another object of the present invention to provide for a tissuetrapping platform which includes some sectionable tissue managementfeatures.

It is another object of the present invention to provide a tissuetrapping platform which includes a method of assuring that the tissuewill be oriented in the desired sectioning plane in the embedding mediaand will be pressed down into the wax embedding material so as to beclose to the sectioning surface.

It is another object of the present invention is to automate the frontend of a biopsy sample analysis procedure by providing a method to placethe harvested tissue appropriately onto the sectionable filter or stageor on a non-sectionable stage prior to tissue processing.

It is another object of the present invention to automate the paraffinembedding process once the tissue has passed through the processor.

It is another object of the present invention to provide a method forautomating the gross in process.

It is another object of the present invention to provide a fine needleaspiration biopsy device which includes a detachable tissue trappingsectionable support means specifically adapted for the needs of specimenprocessing in pathology.

It is another object of the present invention to provide a surgicalbiopsy device which includes a detachable tissue trappingmicrotome-sectionable supporting means specifically adapted for theneeds of specimen processing in pathology.

SUMMARY OF THE INVENTION

These, and other, objects are achieved by providing a multipurposetissue trapping and support means. The tissue trapping means includes atissue trapping and supporting means which can be cleanly sectionedusing a microtome and which is constructed to survive the harsh chemicalenvironment of the tissue preparation process and to be non-distractingduring tissue analysis. For the purposes of this disclosure, a platformassembly includes a cassette frame and either a sectionable immobilizingplatform or a non-sectionable immobilizing platform. By "sectionable,"this disclosure means sectionable in a microtome. The cassette frame isadapted to accept stages or platforms with movable features and isadapted for use in a microtome. The tissue supporting means may be usedin conjunction with a cassette frame, a platform and a cassette, and maybe used to capture tissue samples and to keep them in good conditionduring transportation to the pathology lab and to manage the tissuespecimen during the preparation, wax embedding and sectioning of thetissue.

The tissue trapping means can be used in or in close association withthe harvesting means, such as a Fine Needle Aspiration Device, or thelike, and will support the harvested tissue in a manner that promotesautomation of the handling process, even if the samples are extremelysmall.

Broadly, the invention includes a tissue trapping and supporting meansthat can include a porous member. For ease of discussion, this porousmember will often be referred to as being a filter because it trapscertain material (tissue) while permitting liquid to pass through it.The main purpose of the filter is to trap and hold material, such asharvested tissue samples. The filter is formed so that the tissuesamples received directly from harvesting techniques can be placeddirectly onto the filter and can remain on that filter throughout theentire process, including microtome sectioning and mounting on a slidefor analysis. The filter is microtomable, that is, it can be cleanlysectioned in a mictrotome. In this manner, the handling of the tissuesamples can be entirely carried out in an automated manner because thetissue sample does not have to be handled.

More specifically, the invention includes various tissue trappingplatforms that reduce the amount of sample handling required by eitherthe pathologist or technician and make possible an automated system. Atissue trapping platform includes a filter or stage in a cassette frame.The tissue trapping platforms can have a movable sample surface. Themovable sample surface facilitates sample loading, confers protectionfrom crushing of the tissue samples during the processing steps andallows the sample surface to be pushed into the wax mold for embedding.

One of the platforms described in this disclosure contains a sectionableimmobilization stage which enables the pathologist or technician toorient and fix manipulable tissue samples such as gall bladder, prostatechips or transverse vessel samples. The term "sectionable" as usedherein means the item can be cleanly sectioned into extremely thinsections using a microtome so the layers can be mounted on a slide forfurther analysis. The tissue sample can be stretched or "pinned" into anappropriate orientation to provide for the proper plane of sectioning.This orientation process can take place at initial gross in and only hasto be done one time to ensure appropriate positioning for sectioning.The prior art requires handling of the samples before processing andthen orienting of the samples after tissue processing. The design of thesectionable immobilization stage and cassette frame combination allowsfor the vertical translation of the sample surface, so the samples canbe automatically pressed down into the wax mold base and positionedclose to the sectioning surface of the wax.

Another type of trapping platform contains a sectionable filter designwhich can be used to collect biopsy samples from various biopsycontainers or devices. These sectionable filters are device specific.One such filter has particular application to the handling of FineNeedle Aspiration Biopsy samples. This filter can be manufactured invarious pore sizes. One application for this filter is to include itwith a biopsy sample container. The trapping filter is detachablyretained on the cap of the container and can be removed with asingle-handed motion. It is intended that the filter be placed directlyinto the cassette frame thereby eliminating the step of retrieving thesamples from the sample container. In addition, this particular filteris constructed in such a way as to allow the filter to remain in thecassette frame while it is in the tissue processor. An immobilizationtechnique which permanently affixes the tissue to the stage, filter orplatform could be used with this type of filter prior to tissueprocessing. The filter when removed from the cassette frame can also beplaced directly into the mold for paraffin block preparation withoutfurther manipulation since it can be successfully sectioned onceembedded in the wax.

When biopsy samples are small (1 mm³ or less) it can be hard to locateand position a sample properly in the wax mold and this handling can betime consuming. By retaining the samples on the sectionable filter frominitial collection to the final embedding procedure these problems areavoided.

Another type of sectionable filter platform, also referred to as thesectionable filter cassette, is designed as a screen in a cassette framewith a vertically translatable sample surface. This type of trappingplatform might be coupled with an immobilization technique and thenwould allow for the automatic gross in of Fine Needle Aspiration Biopsysamples as well as mucosal scrapings, endometrial curettes, GI biopsiesscrapings, etc. This sectionable filter cassette could be manufacturedin different pore sizes to accommodate different applications.

A fourth type of trapping platform contains a non-sectionable stagewhich can accommodate large pieces of tissue which do not changeorientation during processing just because of their size. These samplesalso protrude far enough off the surface of the stage so that onceembedded in wax, enough sample is available in the microtomed sectionsso that the non-sectionable stage itself never interferes with themicrotome blade.

The non-sectionable platform with its movable sample surface can remainin a cassette frame through tissue processing, wax embedding andmicrotome sectioning.

The invention includes a means to immobilize the tissue samples on afilter or stage to reduce the number of manipulations required and toenable the automation of the whole histologic section preparationprocess. The immobilization technique does not alter the tissuecomposition in any way, nor does it interfere with the normalinteractions of the tissue and the processor and wax embedder as well asthe appearance of the final section. Immobilization techniques, from thevery simple to more complex are disclosed hereinafter.

In addition, one element of the present invention provides a noveltissue separation system and allows for the recovery of tissue samplesfrom the effluent in a surgical suction device. The combined features ofthis invention reduces the transfer and handling requirements of thesamples throughout the entire process.

Additionally, there is provided a means to immobilize the tissue on aplatform which can then be passed through the tissue processor and waxembedder. The prior art requires the histotechnologist to spend a majorproportion of a work day removing tissue from cassettes after processingand orienting them in the wax mold. The present invention discloses anovel method for eliminating these steps by automating this process.

The present invention provides a reduction in handling by immobilizingthe tissue onto or along with a platform that can travel through theentire tissue preparation and mounting process. The immobilization canbe mechanical whereby the tissue is hooked or pinned or otherwisemechanically bound to the platform. Alternatively, the immobilizationcan take on a much more active roll such as adhesives, coatings, gels orcovering materials. The immobilization also permits automation of theentire process. By fixing the tissue to a platform that can be machinemanipulated, the tissue can be moved and oriented through use of machinecomponents that would otherwise crush or be unable to manipulate tissuesamples. By further making the platforms of a material that can beembedded in the final wax process with no ill effects on the sectioningprocess or to the diagnostic pathological review of the stained tissue,the cycle can be completed with labor savings and accuracy of tissuespecimen preparation.

Automation of the histologic section preparation process is asignificant means of consolidating manpower requirements in thehistology laboratory. In today's hospitals there are consolidationefforts underway to reduce or combine services of area health careproviders. In addition, mergers and takeovers have forced some histologylabs to go to extreme measures to keep up with the demand for processedand sectioned histologic slides. In the prior art, one of the mosttime-consuming tasks in the laboratory is the manual handling of biopsysamples. By reducing the handling requirements and redundant stepssignificant reductions in labor-related costs can be achieved with thisinvention. The present invention includes means and methods to manuallyload or automatically dispense specimens, automatically gross inspecimens, automatically immobilize specimens and automatically waxembed specimens. Use of any of these automated procedures substantiallyimproves the work flow in the histology laboratory and potentiallyprovides the pathologists with their sections for review in a moretimely and efficient manner.

The tissue trapping platforms of the present invention provide a surfaceto which the tissue will become attached at or before gross-in. Meansare disclosed for immobilizing the tissue sample to the filter or stageof the platform prior to introducing it into the processor. The tissueremains attached to the platform through the tissue processor withouteffect on the tissue or processor. This further allows that once throughthe processor the platform and tissue could be handled by mechanicalmeans through the wax embedding procedure and does not necessarilyrequire further manipulation by a technician.

A description of the process with the biopsy container system withintegral sectionable filter will now be presented. The tissue is placedor deposited on the sectionable filter at the time of harvest in thesurgical setting. The sectionable filter and tissue are then immersed ina fixative solution for transport to the pathology lab. Once in the labthe pathologist or clinician removes the sectionable filter from thecontainer. The tissue is trapped on the sectionable filter so there isno need to probe around inside the container looking for tissueparticles. The sectionable filter and specimen are grossed in (describedfor record) and placed in a filter cassette frame. If necessary, at thispoint a tissue immobilization technique can be applied in order to affixthe tissue to the sectionable filter.

The sectionable filter is constructed to survive the harsh chemicalenvironment of the processor. After the cassette has emerged from theprocessor, the sectionable filter/specimen is placed in the embeddingmold, tissue side down. Since the sectionable filter of the presentinvention has been specially formulated from a material that allows itto be sectioned in the microtome, the sectionable filter itself becomesembedded in the wax along with the tissue specimen. This eliminates thefurther step of finding and individually placing each tissue fragment inthe embedding waxing mold. After the sectionable filter is placed in themold, the mold is filled with molten paraffin. When chilled, theparaffin with embedded specimen and sectionable filter (paraffin block)is removed from the mold and is ready for sectioning to make histologicslides.

Still further, because the filter eliminates the need to manually handlea tissue sample, the automated process could also include an automatedgross-in station. In some cases where specific tissue orientation is notcritical, an additional automated step can empty the contents of abiopsy container onto a sectionable platform, depositing the largersamples on the filter surface of the platform. This is applicable tosamples from Fine Needle Aspiration Biopsy and GI biopsies, inparticular. Upon arrival at the histology lab the sample containers areplaced into the automated gross-in station where the machine removes thelid of the container and decants the fluid containing the samples onto asectionable filter cassette (assuming a sectionable filter does not comewith the container as disclosed herein). This process will work well forsamples such as GI biopsy that do not need to be oriented in any specialway for the section.

A surgical pathology accession number, unique to each specimen, isobtained when the specimen is accessioned into the laboratory's anatomicpathology computer system. A barcode can be generated at this time andplaced on the specimen container thus uniquely identifying the specimenwith its accession number. By interfacing an automated computer system,the surgical pathology accession number can be printed on each specimencassette and video image. The number can be human readable and/orcomputer readable. The samples that are trapped on the sectionablefilter are then recorded with a single digital image or infrared orother scan which could have a 1 mm (or other scale) reticule grid infront of the lens to aid in sizing the tissue pieces. The image,surgical accession number, date and other pertinent information arestored on a write optical computer drive or other magnetic media forarchive purposes. Once scanned, the platform with sample is fed into theimmobilization device.

This invention combines elements not previously known in the art tosolve long-standing issues and addresses new innovations in thehistology processing area, as well as other elements that have not beenaddressed to date. This invention provides for a tissue managementsystem that may be implemented in many ways. Several unique tissuetrapping platforms as well as several surgical biopsy devices aredisclosed.

The invention also provides a means and method to more effectivelyobtain biopsy samples via, for example, Fine Needle Aspirationtechniques, along with improvements to the actual cutting surface of theneedle used to harvest the samples as well as the containers used tostore tissue samples.

BRIEF DESCRIPTION OF THE DRAWING FIGURES

FIG. 1 is a filter cassette or stage.

FIG. 2 is another form of filter cassette or stage.

FIG. 3 is a tissue sample mounting means.

FIG. 4 is a sectionable immobilization stage.

FIG. 5 is a sectionable immobilization stage assembled in a stagecassette frame.

FIG. 6 is a non-sectionable stage with biopsy samples thereon.

FIG. 7 is a non-sectionable stage in a wax mold cavity.

FIG. 8 shows the steps in an automated process of handling tissuesamples according to the present invention.

FIG. 9 is a filter stage with a processing number thereon.

FIG. 10 is an automated machine for processing tissue samples fromgross-in to final slide preparation.

FIG. 11 illustrates the processing of an immobilizing platform.

FIG. 12 shows a stage in a wax molding machine.

FIG. 13 shows tissue immobilized on a platform.

FIG. 14 shows the immobilized tissue being placed in a mold for the waxprocess.

FIG. 15 shows a cassette with a processing number thereon.

FIG. 16 shows steps in the operating sequence for a pair of mold bases.

FIG. 17 shows a tissue immobilizing step.

FIG. 18 shows the tissue immobilizing step of FIG. 17 with immobilizingmeans in place.

FIG. 19 shows a container having a filter.

FIG. 20 is an exploded perspective view of the container shown in FIG.19.

FIG. 21 illustrates the use of a tissue sample container.

FIG. 22 illustrates another step in the use of the tissue samplecontainer.

FIG. 23 shows a tissue collection device.

FIG. 24 shows a tissue collection device.

FIG. 25 shows a Fine Needle Aspiration device.

FIG. 26 shows a prior art needle.

FIG. 27 shows a needle embodying the present invention.

FIG. 28 shows the prior art needle removing a tissue sample.

FIG. 29 shows the needle of the present invention.

FIG. 30 is a view taken along line 30--30 of FIG. 29.

FIG. 31 shows a needle using the teaching of the present invention.

FIG. 31A shows a prior art needle.

FIG. 32 shows a needle using the teaching of the present invention.

FIG. 33 shows another view of the needle in FIG. 32.

FIG. 34 shows a tissue harvesting step in the process of the presentinvention.

FIG. 35 shows a step of storing a harvested tissue sample in acontainer.

FIG. 36 shows another step of storing a harvested sample.

FIG. 37 shows a step in the process of storing and handling a harvestedsample.

FIG. 38 is a flow chart showing the process of harvesting and handling atissue sample according to the teaching of the present invention.

FIG. 39 is a perspective view of a laboratory device using thesectionable filter of the present invention.

FIG. 40 is a sectional view of the laboratory device shown in FIG. 39.

FIG. 41 shows a tissue supporting means and tissue embedded in a finalassembly.

FIG. 42 shows a platform, such as shown in FIG. 3, embedded in wax.

FIG. 43 shows a microtome device slicing a wax embedded specimen andtissue supporting means.

FIG. 44 shows another view of the microtome slicing a wax embeddedspecimen and tissue supporting means.

FIG. 45 shows a slide mounted tissue/tissue supporting means/wax whichhave all been sliced in a microtome.

FIG. 46 shows a slide mounted tissue/tissue supporting means/wax whichhave all been sliced in a microtome.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT OF THE INVENTION

Tissue Trapping Platforms

A platform includes a filter or stage assembled in a filter cassetteframe or a stage cassette frame. FIGS. 1 and 2 show platform assemblieswith interchangeable microtome sectionable tissue trapping filters,sectionable immobilizing stages or non-sectionable immobilizing stagesand cassette frames.

Microtome Sectionable Tissue Supporting Means

FIG. 1 shows a cassette frame 10 with a cylindrical interior frame 12which is designed to accept microtome sectionable tissue supportingmeans, such as filters A' and A'' each of which can be porous and formsa tissue supporting means 14 surrounded by a collar 16. As discussedabove, the term "filter" will be used for the tissue supporting meansbecause, in one form of the tissue supporting means, fluid can passthrough the tissue supporting means while tissue samples are retained onthe supporting means in the manner of a filter. Tissue supporting means14 supports tissue samples during tissue processing, embedding andmicrotomy and can include sectionable filters which can be located insurgical biopsy instruments, biopsy containers, or the like, and can beintegral with the intrument or container, or could be used in anautomated biopsy sample dispensing system, with effluent passingtherethrough as will be understood from the ensuing discussion. Themeans for supporting biopsy samples further includes a collar 205surrounding the tissue supporting means and means, such as projections20 on the collar, for connecting the tissue supporting means to theframe 10 via grooves 18. The shape of the tissue supporting means isshown as circular, but could be other shapes as well without departingfrom the scope of the present disclosure. Filter A'or A'' is movablewith respect to the frame 10 and has a means for moving the filterelement into multiple positions relative to the frame, with this meansincluding filter detent grooves 18 internal to the central element 12and projections 20 on collar 16 which mate with the grooves and whichextend radially outward from the periphery of the sectionable filters.There can be several grooves which are spaced apart from each other inthe frame along the longitudinal dimension of the frame. Projections 20can be moved from one groove to another to allow the sectionable filter,and more specifically the sample surface, to be movable and to bepositioned at various heights with respect to the cassette frame ends 13and 13'. As will be understood from the teaching of this disclosure, thevarious heights allow access to the sample surface of the sectionablefilter for more convenient loading and also offer protection fromabrasion or dislocation of the tissue on the sample surface duringtissue processing. The translatable height feature of the cassette framealso allows the sample surface of the filter to be positioned deep intothe mold cavity for wax embedding.

Sectionable filter A' shows a fine 1/4 mm filter grid and sectionablefilter A'' shows a 1 mm filter grid. Preferred pore sizes are 1 mm, 1/4mm and 180 microns to 200 microns for use with FNAB. However, other poresizes can be used based on the teaching of the present disclosure aswill occur to those skilled in the art. The sectionable filter grid canbe manufactured in many other sizes as will be understood by thoseskilled in the art based on the teaching of this disclosure.

In general, the filter is one form of a tissue supporting means used inan overall means for supporting histologic tissue biopsy samples. Ingeneral, the overall means comprises a microtome sectionable tissuesupporting means such as filters A' and A'' for supporting tissuesamples during tissue processing, embedding and microtomy including ameans for permitting the tissue supporting means to be successfullysectioned in a microtome. Successful microtome sectioning means, as usedherein, sliced in a microtome without damaging the microtome or thetissue, or without tearing or cleaving the tissue or the tissue support.The tissue supporting means includes a means for resisting histologicalstains, a means for resisting degredation from solvents and chemicalsused to fix, proces and stain the tissue and a means for maintainingsaid tissue supporting means non-distracting during tissue processingand slide preparation. As used herein, the term "degradation" is definedto mean softening, discoloring or any kind of unfitness for use in allprocesses associated with the analysis of the tissue.

The sectionable filter or stage is made from a special low densitythermoplastic which is molded into a porous filter or screen. The filteris specially selected to resist the chemical and heat environments inthe tissue preparation processor. At the same time the material must beof similar density to both the tissue and the paraffin embeddingmaterials. It must further be able to be sectioned using a standardlaboratory microtome (microtomy) without dulling or nicking the blade.The material must section just as if it were part of the wax withouttearing or cleaving. If the filter material tears during microtomesectioning, it may destroy the fragile tissue section. The material mustalso not stain when the tissue is prepared with various histologicstains. It should not become soft, discolored or dissolved in thesolvents and chemicals used to stain the tissue. Still further thematerial must appear non-distracting, such as window clear, in thesection so as not to distract or confuse the pathologist duringmicroscopic examination. As used herein, the term "non-distracting"means that the material will be readily identifiable as being filtermaterial as opposed to tissue when viewed during analysis of the tissuespecimen. Thus, a "non-distracting" material will not be confused withtissue being analyzed during tissue analysis. The preferred form of anon-distracting material appears window clear or at least translucentwhen viewed during tissue analysis; whereas, the tissue has a color orappearance that is readily identifiable as being tissue. One suchmaterial is a low density polyethylene homopolymer such as QuantumChemical Co. Petrothene® NA 601-04.

Since the sectionable filter will be used to separate small tissueparticles from suspended liquids it may be necessary to modify thesurface tension or wetting characteristics of the plastic to allow thefluid to pass rapidly through the filter screen while retaining samples.Surface treatments such as Plasma etching, Corona Discharge, Ion beam,Hydrogels, or Photolink(TM) surface modifications can be used. Thesesurface treatments may also be used to attract or retain tissue on anyof the filters or stages. As will occur to those skilled in the art,there may be a need to have an affinity coating to attract mucosaltissue fragments as an example.

Stage Cassette Configurations

FIG. 2 depicts a stage cassette frame 10' in which both sectionable andnon-sectionable tissue trapping stages can be inserted. It is arectangular version of the cassette frame 10 shown in FIG. 1. Stagedetent grooves 18' are positioned inside the periphery of the cassetteframe and projections 20' on the stages mate to the stage groovesallowing for various vertical positions of the sample surface to beestablished with respect to the stage cassette frame as above discussed.

Sectionable Filter Cassette Configuration

In FIG. 2, B''' shows a sectionable filter stage for the cassette frameconfiguration. This type of filter could be used to process small piecesof tissue that arrive in the laboratory in a container with fixative andthe container does not already contain an integral sectionable filter.

Sectionable Immobilization Stage

In FIG. 2, B'' shows a sectionable immobilization stage. FIG. 3 depictsthe sectionable immobilization stage B'' assembled in the stage cassetteframe as a platform and ready for tissue loading. FIG. 3 also shows theinstallation of a long thin biopsy sample into gripping pins 22 on thestage. These allow the pathologist to manually orient the tissue samplesfor sectioning at the point of gross in of the sample. FIG. 3 also showsa tubular tissue section TS such as an artery or a vein being installedover a vertical tissue pin 23. This allows for transverse section of aluminal structure.

Additional hooks 24, pins and gripping elements can be provided on thesestages to allow the pathologist to select the most appropriateimmobilizing method and orientation for each tissue sample. As shown inFIG. 3, the gripping features and actual sample surface of the stageextend above the stage cassette top rim 25 when the stage is positionedin the top most groove. This facilitates tissue loading.

FIG. 4 again shows the sectionable immobilization stage B'' assembled instage cassette frame 10' and turned upside down. In this view thegripping features and sample surface are beneath the stage cassetteframe rim 25, offering protection to the tissue samples from dislocationduring tissue processing. The stage has been adjusted to rest in lowergroove 18'L of the stage cassette frame.

FIG. 5 shows sectionable immobilization stage B' positioned once againinto upper stage groove 18'U of the stage cassette frame, as it was fortissue loading. In this view, however, the sectionable immobilizationstage and frame have been turned upside down and are shown pressed intomold cavity 27 of a wax mold 28 ready for embedding in wax or paraffin29. The tissue samples are thus presented as close as possible to theeventual sectioning surface.

Non-sectionable Stage

In FIG. 2, B' shows a non-sectionable stage on which large tissuesamples can be immobilized. The non-sectionable stage can be used whenthe sample is large enough that it extends well above the sample surfaceof the stage and sections of the wax embedded sample can be made withoutrunning into the actual stage with the microtome. FIG. 6 shows across-section of the non-sectionable stage assembled into a stagecassette frame 10'. The stage has projections which engage the stagedetent grooves to allow for the vertical translation of the stage withinthe cassette frame whereby biopsy sample BS can be oriented and locatedfor proper treatment. Movement of the filter or stage with respect tothe frame is effected by simply pushing the filter or stage with respectto the frame. The filter or stage and the frame are made of flexiblematerials and thus will deform when such pressure is applied. Thisdeformation will permit the projections to pop out of one groove andthen slide until they reach the next groove. At that point, theprojections will pop into that groove.

FIG. 7 shows the non-sectionable stage and cassette frame in theposition for wax embedding in the mold cavity. Here, the tissue sampleis presented as close as possible to the eventual sectioning surface. Aswill be discussed below, an immobilizing media keeps the tissue in placeon the sample surface of the stage during tissue processing and waxembedding.

The tissue trapping platforms have numerous applications for use.Specific applications of the invention are discussed herein.

These are shown as examples of methods to trap and transport tissuesamples to the histology lab. It should be noted that there may be manymore uses for this technology so as not to limit the tissue trappingplatform concepts and applications to the disclosed applications.

In general, the invention includes a method for preparing biopsy tissuesamples for histological examination comprising: removing a tissuesample from a patient; placing the tissue sample onto a support;immobilizing the tissue sample on the support; subjecting both thesupport and the tissue sample immobilized thereon to a process forreplacing tissue fluid with wax and impregnating the tissue sample withwax, embedding the tissue sample in a wax mold to form a solid block ofwax, using a microtome, slicing the solid block of wax into thin slices;and mounting at least one of the thin slices on a support member forexamination. It is also noted that one form of the invention includes atissue support that can be successfully mictrotomed, while another formof the invention includes a support that is porous. In that case, thetissue support will be embedded with the tissue sample in the wax andboth the sample and the support will be sectioned using a microtome.

The invention also, broadly, includes a tissue analysis automationprocess which includes placing tissue on a machine manipulable supportmeans; immobilizing the tissue on the support means to maintain aselected orientation of the tissue on the support means; and processingthe immobilized tissue along with the support means to replace tissuefluids with wax, as well as a method of conducting analysis of tissuebiopsy samples comprising: harvesting tissue samples from a patient;placing the harvested tissue samples onto a machine manipulable tissuesupport; immobilizing the tissue samples on the tissue support; andprocessing the tissue samples and the tissue support to replace tissuefluids with wax.

Automation of Tissue Processing and Histologic Section Preparation FIGS.8, 9, 11-16 illustrate a method for automating the gross in process, theimmobilization of tissue and the tissue embedding process. Currently,the histotech or pathologist performs all of the preparation proceduressuch as grossing in the samples and placing the tissue into cassettesprior to putting them in the tissue processor. The histotechadditionally performs all of the manipulations required to place thetissue into the molds for paraffin embedding.

Automated dispensing of samples in fixative solution

FIG. 8 depicts a process whereby small biopsy samples are removed fromcontainers and filtered through a sectionable filter. In FIG. 8, Step 1shows a bar code reader 402 which reads a digital bar code 401 off theside of biopsy container 404. This bar code number is matched to alaboratory accession number which is used to track the tissue samplesthrough the processor and the wax embedding station.

Step 1 of FIG. 8 shows the biopsy container 404 placed in containergripper 403. Bar code 401 is read and recorded by reader 402 whichcommunicates with CPU 417 via central data bus 412. Bar codes and barcode readers, as well as the associated computer equipment and softwarethat are used for this process are known to those skilled in the art.Therefore, based on the teaching of this disclosure, those skilledartisans will be able to select this equipment and software. Therefore,such equipment and software will not be further discussed.

In Step 2, cap gripper 406 powered by cap removal servo 407 removes anddiscards the standard biopsy container cap 405. Cap grippers, as well asthe other mechanical equipment necessary to carry out the steps and topractice the invention disclosed herein, will be known to those skilledin the art based on the teaching of this disclosure. Accordingly,details of such equipment will not be presented.

In Step 3, the container is tilted and the contents are dispensed into afunnel stage 413 directing the contents first through sectionable filter400. Again, as discussed above, such elements are known to those skilledin the art. As depicted, this is a sectionable filter but couldalternatively configured as a sectionable filter cassette. The filtrateis normally discarded as waste, however, the filtrate can be directedinto a cytospin container 414. When a cytology test has been ordered, itis indicated by the technologist or pathologist by placing a specialcytospin indicator 427 on the biopsy container. Bar-code reader 402notes the indicator and directs the appropriate pore size sectionablefilter to be automatically installed along with cytospin container 414.The cytospin container is automatically labeled with the biopsy sample'sunique accession number with printer head 424 (machine and/or humanreadable). The funnel 413 could be a single use device which is disposedof after use to prevent cross contamination of specimens.

Whether or not a cytospin container is used, a rinse cycle is initiatedafter the biopsy container is emptied. Washing wand 409 dispenses rinsesolution from reservoir 411 controlled by rinse valve 410. The rinsesolution will clean out the biopsy container of any particulate thatwill then, if large enough, be trapped on sectionable filter 400. Thesmaller cellular components of the biopsy flow through filter 400 andare either discarded or captured in cytospin container 414 for furtherprocessing.

Automated Gross in of samples

In Step 4, filter stage 400 moves into a gross in station where an imagecapturing device such as a digital camera or video camera 415 records animage 416 of the tissue samples on sectionable filter 400. A 1 mm (orother gradation) grid reticule on the camera lens may be used for a sizecalibration. Traditionally, the number and size of tissue samples in acassette are described at gross in for future reference. With thepresent invention, the digital picture contains a record of thisinformation, that may be printed on the surgical pathology report, ormay be accessed sometime in the future if questions arise. The digitalpicture is displayed for the histotech to verify that a record has beencreated for the particular biopsy sample. The information is digitallycompressed by CPU 417 and stored on an optical disk or other datastorage media 418. In addition, given the information in the digitalimage, the processor can determine if too much tissue is present on agiven platform and reject it for further treatment by the histotech.Additional scanners such as infra red could be used as a diagnosis tool.

In addition, the digital image can be used to transcribe an appropriategross description of the surgical pathology report of the specimen. Thedigitized image is analyzed by CPU 417 to determine the number and sizerange of the pieces of tissue in the specimen. This information ispassed through an interface to the laboratory anatomic pathologycomputer system. Through appropriate programming, such as macros similarto ones which are presently in use in the word processing systems ofmost laboratory systems, the system transcribes an appropriate grossdescription of each specimen for incorporation into the surgicalpathology report.

CPU 417 controls all of the stations in steps 1 through 5 and recordsall events, tracking numbers on digital storage media 418. An accessionor tracking number 423 (see FIG. 9) is printed (machine and/or humanreadable) on the cassette frame to identify the samples. This numberrelates to the bar code number from the biopsy container if theautomated decanting process was used; or to a sequential log numberwhich is also printed on a label and presented to the technician toattach to a requisition form with specific information about the originof the sample; or the information would be tied to the computerized logbook in the histology lab.

Immobilization of tissue on platform

The tissue immobilization process on the filter or stage will now bediscussed. Both manual and automated immobilization techniques have beenproposed. The pathologist or technician is able to properly orientsamples as required for sectioning by placing the samples on anappropriate tissue trapping platform just prior to gross in. Theimmobilizing process maintains the pathologist-specified orientation ofthe tissue throughout the histology preparation process. No furthertreatment is required for samples placed on a sectionable immobilizingplatform since its gripping features act to hold the tissue in placethroughout the histology preparation process. The tissue on anon-sectionable platform or sectionable filter cassette may requirefurther immobilization treatment.

Tissue immobilizing means can be included on the tissue supporting meanswhereby tissue samples are quickly immobilized on contact with thetissue support. For instance, cyanoacrylate adhesive works well to bondlarger tissue samples to the non-sectionable stages. The adhesive curesquickly and bonds the tissue securely and further does not break down inthe processor fluids. Additional adhesive-like substances can be coatedon the surface of the filter or stage making a "dry adhesive" which canbe activated by the moisture in the tissue sample. Additionallyultraviolet curing dry adhesives can be used; the adhesives are drycoated and do not become "activated" until catalyzed with ultravioletlight. Still further, coatings with protein affinity can be depositedupon the filter or stage whereby contact with any protein containingmaterial will catalyze the adhesive. Other tissue immobilizingtechniques and methods can include the techniques of Dry Net, ballisticparticle deposition and use of adhesives are disclosed as well as othermethods ranging from simple to complex.

After gross in, the filter stage is move by appropriate machinery aswill be understood by those skilled in the art, into Step 5 of FIG. 8which is an immobilization step. If the sample has been placed onto asectionable filter, a sectionable filter cassette, or a non-sectionablestage, immobilization is desirable to keep the tissue samples in placewhile in the fluid medium of the tissue processor. In this embodimentthe immobilization device is shown as a ballistic particle head 419 fedby a heated pressurized reservoir 420 of material such as low densitypolyethylene. The ballistic particle head 419 is on an x-y gantry 424which enables the deposition of a fine web-like netting to be createdover top of the samples. Using information from the digital image takenin Step 4, an "intelligent net" could be created specifically capturingpieces of tissue rather than covering the whole filter or stage surface.

Although the preferred embodiment is shown as ballistic particledeposition of material, many other ways could accomplish the same resultsuch as the thermal bonding of a net material over the tissue (Dry Net,see FIGS. 17 and 18); or by spraying a thin glue-like substance over thetissue and filter or stage; or by spraying a thin glue-like substance onthe sample surface of the filter or stage prior to tissue loading; or byspraying a thin layer of agar or other gel over the tissue and filter orstage; or by using a bio-affinity coating that would allow the tissue tobind to the surface of the filter or stage after exposure to anultraviolet cure period or without the ultraviolet cure; or by using anultraviolet cure adhesive coating on the filter or stage surface; or byusing a coating of albumin or L-Lysine or some other sticky protein onthe surface of the filter or stage. Such alternatives will occur tothose skilled in the art based on the teaching of the presentdisclosure.

In the Dry Net technique the tissue on a platform is then placed into animmobilizing fixture seen in FIG. 17. The immobilization fixture bringsthe platform underneath a polyethylene net 30 which is fed by rollers 31from a feed reel 32 to a take-up reel 33 toward a transfer base 34 onwhich the cassette is supported. The web is moved by a stepping motor 35connected to the rollers. The platform is positioned underneath the netand a bonding head 36 is brought down from above to ultrasonically orheat weld the net to the periphery of the platform (FIG. 18). This trapsthe tissue between the sample surface of the filter or stage and thenet. The net is preferably made from the same sectionable material asthe filter or stage and is preferably porous so that the tissueprocessing fluids and the wax can penetrate around the tissue.

Two methods for use of a wet adhesive process to immobilize the tissueon a filter or stage are disclosed whereby either an adhesive is sprayedonto the sample surface of the filter or stage prior to loading with thetissue sample; or the adhesive is applied after the tissue has beenplaced on the filter or stage. In order to be effective, the lattermethod requires the adhesive to wick underneath the edges of the tissueand thereby hold down the tissue throughout processing. Adhesives suchas cyanoacrylates are well suited for this application since moisturesets off the rapid curing process. Tests have shown that thecyanoacrylate--tissue bond is impervious to the chemical and temperatureenvironments of the tissue processor and the wax embedder. It does notinterfere with the sectioning or staining of the samples nor does itinterfere with the tissue histology.

Any substance with does not interfere with the histologic samplepreparation, as described above, can be used to immobilize and affix thetissue to the platform. The immobilization process depicted in FIG. 8uses ballistic particle deposition in which small particles of moltenplastic are ejected from a nozzle towards the filter or stage andtissue. The ballistic particle technology is currently in use in therapid prototyping process whereby plastic models are constructed fromthree dimensional CAD files. Since those skilled in the art willunderstand this technology, it will not be further discussed.

If the tissue sample has been loaded onto a sectionable immobilizingstage which does not require an extra process to secure the tissue toits surface, a machine readable code on the platform could identify theplatform type and allow for this specific type to bypass theimmobilization step and continue on to the tissue processor.

FIG. 9 shows immobilized tissue 422 with immobilizing material 421retaining the tissue on a sectionable filter. The tissue will not bedislodged from the sample surface during further processing.

Once the tissue is immobilized on the filter or stage, the platform canbe placed in a standard storage rack or automatically introduced intothe tissue processor. If the filter or stage is not already in aplatform configuration, it will be automatically placed into theappropriate four sided cassette frame before progressing to the tissueprocessor.

Automated Wax Embedding Process

In general, the automated process prepares the tissue sample forembedding in wax, and embeds the tissue sample in wax. Then, the tissuesample can be sliced into thin slices using a microtome and at least oneof the slices mounted for microscopic examination. Broadly, the methodof preparing biopsy tissue samples for histological examinationcomprises: removing a tissue sample from a patient; storing the tissuesample in a container; dispensing the contents of the container onto asupport; immobilizing the tissue sample on the support; subjecting boththe support and the tissue sample immobilized thereon to a process forreplacing tissue fluid with wax and impregnating the tissue sample withwax, and embedding the tissue sample in a wax mold to form a solid blockof wax. As above discussed, one form of the invention includes a poroustissue support while another form of the invention includes a tissuesupport that can be successfully sectioned by a microtome. A microtomeis then used to slice the solid block of wax into thin sections whichcan be used for further examination. If the tissue support ismicrotomable, it, or part of it, can also be embedded in the wax block.

More specifically as shown in FIG. 11, as platforms 426 emerge from thetissue processor they can be stored in a rack 450 for batch processingor sent directly to the automated wax embedding station. FIGS. 11 and 12illustrate an automated wax embedding station. Flip down fixture 452 atthe end of the storage rack 450 includes a means to transfer and orientthe platform with the tissue face down in the wax mold. When theplatform comes into position in the rack, sensors note this and activateactuator 451. Actuator 451 includes a cylinder and is operated by amotor (not shown) to rotate flip down fixture 452 into the horizontalorientation to enable the pick and place head 457 to access the upsidedown platform. An ear 33 is rotatably connected to rack 450 tofacilitate this movement. Pick and place head 457 has three functions: alongitudinal function shown as an arrow 461; a vertical picking head 454and a setting head 453. Vertical picking head 454 can move vertically onstage 456 by means of a motor (not shown). Actuator 455 moves settinghead 453 vertically via a motor (not shown). Mold base 432 is one of apair of mold bases used in this machine. Each mold base has a mold subbase 429 which houses molding cavity 434. Additionally, the system hastwo paraffin dispensing stations 460 which include hot molten paraffin428, heated reservoir 458 and dispensing tip 459. Mold base 432 can beactuated to move in a linear fashion from left to right. Movement of theelements of the wax embedding processor are controlled by motors whichare, in turn, controlled by computers. These control elements are notshown as one skilled in the art will understand what is required ofthese control elements based on the teaching of this disclosure.

FIG. 16 depicts the operating sequence for the pair of mold bases.Referring to FIGS. 11 and 16, it can be understood that in Step 1₁₆, theavailable mold base (labeled A₁₆) is moved over to the paraffindispensing station 460 where in Step 2₁₆ a small quantity of paraffin isdispensed into mold cavity 434 prior to the platform being positioned inthe mold. This provides a thin rapidly cooling layer of paraffin in thebottom of the mold, for the tissue and filter or stage surface to be setinto. In Step 3₁₆ the mold base then traverses under flip down fixture452. The pick and place head 457 comes down onto the platform, grips itand sets it into the paraffin layer in the mold cavity. FIGS. 13-15depict a sectionable filter 425 that has been dropped into a filtercassette frame 426 and is shown flipped over, ready for embedding in thewax mold form. The process is identical for the other platformconfigurations.

In FIG. 13, immobilized tissue 422 is protected by cassette frame 426while in the tissue processor since the sample surface is verticallycentered within the cassette. Setting head 454 (FIG. 11) applies andmaintains pressure on the sectionable platform which translatesvertically downward within the cassette frame into mold cavity 434. Thisdownward vertical translation is depicted in FIG. 14 and can be seen bycomparing FIGS. 13 and 14. This insures that the tissue samples are setin the bottommost position in the mold cavity and therefore, whensectioning, the microtome will have easy access to the tissue sample. Ascan be seen in FIG. 12, to facilitate the setting of the paraffin layer,mold sub base 429 is cooled via cooling channels 430 which surround thecavity 434. The cooling channels are connected to tubes which lead to aseparate chilling unit for circulating cooling fluid to maintain atemperature at the mold sub base of approximately -7° C.

Once the platform and tissue samples are set in the pre-fill layer ofparaffin, the pick and place head is raised and the mold base is againtranslated laterally to the paraffin dispensing station 460. In Step 4₁₆the mold is automatically filled to the final level. Mold base 432dwells at this station for a period of time (Step 5₁₆) post fillingduring which time the sub base is chilled to set the newly addedparaffin.

In Step 1₁₆ again mold base 432 translates back to the center positionwhere the pick and place head 457 comes down and removes the platformfrom mold sub base 429. In order to facilitate easy removal of thehardened paraffin block and attached platform 426 from the mold subbase, the mold sub base is pivotally mounted on the mold base withactuating mechanisms 431 that are operated and controlled by computercontrolled motors (not shown). Mold sub base 429 is preferably made of aflexible material such as urethane, which allows the mold to be flexed,popping the hardened paraffin block out as shown in FIG. 12.Additionally, the paraffin will not stick to the urethane material. Withthe paraffin block removed, the cycle for one mold base is complete.FIG. 16 depicts how two mold bases (A₁₆ and B₁₆), one pick and placehead and two paraffin dispensing stations can be used to improveefficiency of the embedding process. A₁₆ and B₁₆ mold bases alternatestations in an efficient work flow pattern.

The automated process will allow the completed paraffin block to betransferred directly to the microtomy station where it is sectioned,applied to a glass slide and stained.

Any or all of the above described automated stations could be configuredinto a package to best meet the needs of a particular laboratory.Automation of every step would not be a requirement.

FIG. 10 shows a finished product design of a fully automated system. Itis comprised of automated processing stations for each of the majorsteps for histology sample preparation; each could be used individuallyor as a completely automated system. There are five automated stationsshown:

1₁₀. automated sample dispensing and platform selection

2₁₀. printing and video gross in unit

3₁₀. immobilization

4₁₀. tissue processing (prior art technology) and

5₁₀. automated wax embedding.

Additionally, biopsy container storage means 6₁₀ is located adjacent toa gross in location.

In the first station, automated sample dispensing and platformselection, biopsy sample containers C₁₀ are stored in a rack awaitingautomated dispensing. Blank platforms and cytospin containers are storedin an area 7₁₀. Sample containers are brought into the automatedprocessing system. A bar code reader deciphers the machine readable codeon the container, which indicates whether a cytospin container isrequired for this particular sample. The sample container isautomatically opened and the contents are decanted onto a sectionablefilter. If required, the eluate is collected in a cytospin container forfurther processing. There is also a single container entry tray whichcan be used to accommodate a sample which needs to be processedimmediately; samples entered there are given priority over samples thatmay already be in storage.

In the second station a printer head prints a laboratory accessionnumber A₁₀ from the laboratory log records onto the cassette frame andcytospin container if one was required. The platform is moved fordigital or video gross in and display on screen 8₁₀ by video camera 9₁₀and a single digital or video image is recorded of the tissue samples onthe platform, capturing the identifying accession number as well. Amanual loader 10₁₀ can also be used.

A single entry tray 11₁₀ can be provided at this station as well toallow entry of platforms which are loaded manually such as thesectionable or non-sectionable stage platforms or a sectionable filtercassette that has been manually prepared. The printing and video grossin functions are performed on these samples as well.

The platforms are then moved individually into the third station 3₁₀ forimmobilization of the tissue samples. The immobilizing technique isapplied to the tissue and filter or stage and current sample number isdisplayed on screen 14₁₀.

The platforms with immobilized tissue samples are transferred into aholding tank 15₁₀ for batch processing or are sent directly into thetissue processor for continuous processing.

From the tissue processor, the platforms move into the automated waxembedding station (station 4₁₀). They may also be held in storage thereand processed in a batch, if required. The automated wax embeddingsystem described in FIGS. 11, 12, and 16 is housed within this unit.

Finished paraffin block storage trays are provided in which the systemwill store finished embedded platforms awaiting sectioning.

FIG. 38 is a flow diagram depicting the process flow of automatedhistology sample preparation. Process 101 is tissue harvest. Tissueharvest can be accomplished with any surgical device such as fine needlebiopsy aspiration or a surgical biopsy sample device. If appropriate, atissue sample container can be fed into the automated sample dispensingdevice. A bar code reader can read any machine readable information onsample containers or devices which will then match up with thelaboratory accession number from data base 115. If a cytology sample hasbeen ordered, it is noted in box 112 and a cytospin container isautomatically provided to collect the eluate.

In Step 1 other samples are manually loaded onto the appropriate tissuetrapping platforms. The printing station 113 then prints the accessionnumber assigned by the automation CPU form accession log data base 115.This is printed both onto the cassette frame and the cytospin containerif one was required. The cytospin container is exited from the system inStep 2.

Automated gross in 117 is performed and the information is stored onstorage media 118. Decision process 119 determines whether furtherimmobilization of the sample is required. For example, sectionableimmobilization stages which have been manually prepared will not requireapplication of additional immobilization techniques. A machine readablefeature on the stage determines whether the immobilization stationshould be bypassed or not. If immobilizing is required, the platform istreated with the appropriate technique at box 120.

After immobilization, the platforms are held in batch in a processholding tank awaiting tissue processing or are sent continuously throughthe processor. Step 4 is tissue processing which relies on standardprior art technology.

After processing another decision block determines whether the platformswill be held for batch wax embedding or will be embedded as availablefrom the processor. Step 5, box 125 is the automated wax embeddingprocess.

Tissue specimen container with integral sectionable filter

The object of the container embodiment (shown in FIGS. 19, 20, 21 and22) is to provide a simple and easy convenient way to place tissuesamples on the sectionable filter; to detachably hold the sectionablefilter in place on the container while depositing the samples, to retainthe samples on the sectionable filter, to keep the sample wetted withfixative and to provide a convenient way to remove the sectionablefilter and sample from the container without leaving behind any usefulsamples.

In general, one form of a tissue sample container 200 is shown in FIGS.19-22 and includes: a means for supporting histologic tissue biopsysamples which includes a tissue supporting means for supporting tissuesamples during tissue processing, embedding and micotomy and includingmeans for permitting said tissue supporting means to be sucessfullysectioned in a microtome, means for resisting histological stains, meansfor resisting degredation from solvents and chemicals used to processand stain the tissue, and means for maintaining said tissue supportingmeans non-distracting during tissue preparation and slide preparation.As above discussed, one form of the invention includes a tissuesupporting means that can be successfully sectioned in a microtome,while another form of the invention includes a tissue supporting meansthat is porous. Specifically, container 200 includes a body 201, asectionable filter 202, a cap 203, and a gasket 204. An injection site202' is located adjacent to the filter whereby samples can be placed onthe filter. Container body 201 is configured as a wide mouth vessel withconcentric flexible release fingers 205 projecting from bottom internalsurface 206. These fingers are adapted to detachably engage and retainsectionable filter 202. A small retention ridge 207 on an extending lipof each finger engages the sectionable filter collar or ring 202 tolightly retain the sectionable filter on fingers 205 during tissueplacement and transportation. The sectionable filter ring has acorresponding undercut 208 to engage the retention ridge 207 on eachfinger. The sectionable filter is positioned close to the same height asthe container's outer lip 209. This allows samples to be placed orscraped onto the sectionable filter without reaching down into thecontainer. Further, if the sample is being transferred from a longscraping tool is must be able to lie flat on the sectionable filter totransfer the sample. The height of fingers 205 in the container alsospace the sectionable filter just above the fixative level 210 in thecontainer.

The sectionable filter ring is adapted to have an outer ring 211 andspider ribs 212 (FIGS. 20 and 21) that create a support structure forthe filter or screen. It is envisioned that the sectionable filter willbe injection molded in a single unit. The ring has an outer edge 211'that is larger in diameter than the inner diameter of the ring to act asa deformable sealing lip which will allow it to create a seal to theinside bore of displacement cylinder 215 in the cap. Screen 213 ismolded to provide openings in the 0.006" to 0.008" range. Smaller orlarger openings could be manufactured to accommodate the tissue samplesizes desired. The sectionable filter has a small ring 214 thatprotrudes above the screen surface 213'. Ring 214 allows fluid to bepoured through the screen so that it will and not spill over the edge.It is also utilized as a standoff when the sectionable filter is placedin the wax mold to allow any protruding tissue to stand above the screensurface. This prevents any flattening or distortion of the tissue sampleprior to wax embedding. It also provides a surface for heat-sealing animmobilizing net (Dry Net) over the tissue samples.

Yet another feature of the container is the fixative fluid displacementmeans, such as cylinder 215 on cap 203. The cap has an elongatedcylinder which extends below attachment section 216 which is shown asthreaded, but could take on any of a number of configurations, such as:1/4 turn locking; friction or snap fit. The displacement cylinder 215acts to raise the fixative level 210 above the sectionable filter 202inside the container when the cap 203 is installed. FIG. 19 shows asectional view of the cap and sectionable filter in place raising thelevel of fixative 210B above the screen upper surface 213' helping tokeep the tissue samples wetted during transport. This will help toprevent the samples from becoming dried out and will additionally keepthem confined in an area that will strain the fluid contents through thesectionable filter as the cap is removed and the fluid level dropsinside the container.

To facilitate the removal of the sectionable filter from the container,retention ridges 207 on fingers 205 and grooves 217 are fashioned on theinside diameter of displacement cylinder 215. As shown in FIGS. 20 and21, as the cap is lifted up (FIG. 21) grooves 217 engage outer sealingedge 211 of sectionable filter 202 transferring it from container 201 tocap 203. Fixative level 210 will drop as the displacement ring iswithdrawn from the container straining the tissue fragments through thesectionable filter. Sectionable filter 202 can be removed from the capby placing forceps 218 (FIG. 22) into cutouts 219 in the displacementring and disengaging it from the cap. The sectionable filter with tissuesamples are then placed into either a standard prior art tissue cassette220 for non-automated processing, or into a specialized filter cassetteframe for automated processing as discussed above.

Alternately the displacement cylinder would have no grooves to engagethe sectionable filter. This would be necessary case one wants toinspect the filtered contents before removing the sectionable filterfrom the container. In that case, it is envisioned that the sectionablefilter would reside above the lip of the container to facilitate accessto the edge of the sectionable filter with forceps for easy removal ofthe sectionable filter. The cap would retain the fixative displacementring but would not include the retaining grooves.

Fine Needle Aspiration Biopsy Device.

In general, the invention includes a tissue sample container comprising:a means for supporting histologic tissue biopsy samples which includes atissue supporting means for supporting tissue samples during tissueprocessing and embedding and micotomy including, means for permittingthe tissue supporting means to be successfully sectioned in a microtome,means for resisting histological stains, means for resisting degredationfrom solvents and chemicals used to process and stain the tissue, andmeans for maintaining the tissue supporting means non-distracting duringtissue preparation and slide preparation. As before, one form of theinvention includes the tissue supporting means being porous as well.

FIG. 25 depicts a fine needle aspiration device 501, with an integraltissue trapping sectionable filter 502. The sectionable filter ispositioned within the body of the syringe 503, opposite the proximal endof fine needle 504. This allows the physician to take the sample byprior known procedure but assures that larger tissue samples will beretained by the filter in preparation for histologic cell blockpreparation. Retaining cap 505 is threaded for easy removal. This allowsfor removal of the filter by unscrewing the retaining cap and pushingplunger 506 forward to eject the filter. In addition, the physician canelect to prepare a direct smear on a glass slide by first taking thebiopsy then aspirating any fine cellular particles out onto a glassslide.

In order to provide tissue specimens for histologic exam one must firstobtain sufficient quantity and size from the biopsy. As prior art hasshown many attempts have been made at providing fine needle aspirationbiopsy needle configurations that provide improved sample harvestingproperties. Yet in most cases physicians continue to use standard threebevel grind venipuncture needles such as is shown in FIG. 26, mostlikely due to their low cost and accessibility. However, pathologistshave noted that there is a high incidence of insufficient or poorquality samples obtained by the standard venipuncture needle.

If one looks at a venipuncture needle tip under magnification, it willbe found that the tip has three flat faces 510, 511, 512, two of which510, 511 create the sharp tip and a third 512 which is transverse to theaxis at a very acute angle usually 18-20 degrees. The two tip bevels arevery finely ground and produce exceptionally sharp edges 513, 514 thatpart the tissue on insertion. Third surface 512 is less fine and in facthas one serious flaw that creates problems for the cutting of biopsysamples. Edge 515 which is created from the inside bore and the thirdsurface is not well controlled and most often is found to have beentreated by an abrasive grit blast to de-burr the edge. For venipuncturethis is advantageous since it is not desirable to cut holes in a bloodvessel which would cause trauma and bleeding. But when it is desired totake tissue samples, it produces poor and unpredictable results. Itmight be assumed that just honing the third surface to produce a finesharp edge would produce better results, and while this is partiallytrue, the inventors have discovered that the tissue tends to "tent" uponpassage through tissue. FIG. 28 shows the outer edges of the prior artneedle 516 creating "tent poles" stretching the tissue taught betweenedges 516. This prevents the tissue from contacting internal edge 515even if it is sharp. The present invention overcomes this limitation byincluding a four bevel grind 520 shown in FIG. 27 with areas 521₁, 521₂,521₃, and 521₄ which in effect moves sharp tissue severing edge 522 tothe outside or top of the tent. FIGS. 29 and 30 show the two new edges524 and 526 that are created from the inside bore of the needle wherethey intersect the two new flats. This configuration cuts well andprovides adequate tissue samples for histological exam.

When creating a design for FNAB, it must be kept in mind that althoughstandard venipuncture needles are less than optimal, they areinexpensive. Therefore, it is desirable to make the FNAB needle of thepresent invention inexpensive to manufacture. The four bevel grind isrelatively inexpensive to manufacture. However, it is very aggressiveand cuts on the entry stroke. The entry stroke leads to tissue samplesfrom the path to the target site in addition to the target. Anotherconfiguration allows for sampling on the removal stroke. FIGS. 32 and 33disclose a back-eye 617 which is cut through the needle 619 directlyopposite the bevels. This can be manufactured by drilling or EDMmachining. The eye 617 is cut at a severe angle (18-20 degrees) backtowards the proximal end to produce a sharp cutting edge 618 at theneedle's outside periphery. This needle can be inserted to the properdepth and then stroked in and out while applying suction from thesyringe to harvest the samples. The suction has been shown to increasethe quantity of samples retrieved, so it is believed to bring the tissuein closer approximation to the sharp cutting edge.

In yet another improvement the inventors have discovered that any ledgesor interstices in a syringe will create traps where the tissue samplesmay become lodged and therefore become trapped and not retrieved fromthe device for examination. One such area in the standard needle andsyringe is the luer fitting. A prior art needle NP is shown in FIG. 31Aand has a ledge L formed at the exit of the proximal end of needletubing TP in front of tip T of male luer fitting MP on the syringe. Thisledge often traps small tissue fragments as indicated in FIG. 31A. Theinventors have designed their needles 620 to protrude all the way up thecentral bore 621 of the luer fittings eliminating this tissue trappingledge. This can be understood by comparing FIGS. 31A and 31. As shown inFIG. 31A, the adapeter MP has an entrance/exit location E formed by theintersection of the inner surface SI of sidewall S and the inner surfaceMPI of luer adapeter fitting MP. The inventors have extended tubing TPso that the proximal end thereof lies in a place containing intersectionE.

In still another way to implement the sectionable filter technologythere is provided an improved tissue harvesting fine needle such as theones described above, but which deletes the sectionable filter in thesyringe body. This then allows the physician to use a better method ofensuring complete capture of the harvested biopsy samples. Since manytimes the physician will request cell cytology and cell blockpreparation for histology, it must be assured that all sample materialis collected and preserved in fixative immediately after harvest.

FIGS. 35, 36 and 37 disclose an improved container that provides thisadvantage. Syringe 723 and special needle 722 are used to obtain thesamples of target lesion 724 by prior art techniques (FIG. 34) and arethen used for introducing tissue specimens into a container 725. A fineneedle aspiration device can also be used in place of syringe 723.Syringe needle 722 is then inserted into special container 725 throughan injection port 726 in the cap 727 of the container, here cap 727 ismolded of an elastomeric material which allows for an integral injectionport 726 to be included in the cap. The cap has a metal ring 728 whichimparts a compressive force on the injection site to keep it fromleaking when the needle is removed. Fixative solution 729 is sucked intothe syringe body which flushes harvested biopsy samples 730 into thebore of the syringe. Plunger 731 is depressed and the fixative andsamples are then transferred into sectionable filter container 725. Thisprocedure can be repeated as necessary to dislodge any samples. Theneedle is removed from the injection site and the syringe and needle arediscarded. The biopsy samples can now be transported to the histologylab for preparation. Another feature of the system involves the removingof sectionable filter 732 which strains the fixative 733 solutionthrough sectionable filter 732 leaving larger samples 730 on thesectionable filter for cell block preparation and allowing smaller cells734 to pass through the sectionable filter. These smaller cells can thenbe processed as a cytospin cytologic preparation. Still further, FIG. 37depicts cap, 727' fashioned from an elastomeric material, which can beflexed in the correct way to move retaining legs 735 which hold the edgeof sectionable filter 732, outwardly to release the sectionable filterfrom the cap. This allows the histotechnologist to deposit thesectionable filter and samples into the standard prior art tissuecassette 220 with one hand.

Since this sectionable filter fits into the smallest inner dimension ofthe wax mold form, it is not necessary for this particular platform tohave the vertically translatable sample surface feature of the inventivefilter cassette frame. When the sectionable filter is removed from thecassette frame and placed into the wax mold form, the sample surfacewill be automatically oriented in the sectioning plane close to thesectioning surface of the wax mold.

Surgical Biopsy Devices with sectionable filter

FIG. 23 depicts a surgical biopsy device 800 which uses a tissuesupport, such as the above-discussed sectionable filter to trap andtransport tissue samples from the surgical suite to the pathologylaboratory. Surgical biopsy device 800 includes a device handle 802 anda hollow shaft 804 and biopsy jaws 806 with an integral filter housing808. Biopsy jaws 806 can take the form of any number of biopsy jawconfigurations.

Shaft 804 which connects handle 802 to jaws 806 actuates the biopsy jawsand allows for a hollow central channel to transport the biopsy samplefrom the patient's body at the biopsy jaws to the filter surface whereit is trapped. Filter assembly 810 is shown in FIG. 24 and containssectionable filter 812. The filter assembly is installed in a filterhousing 814 which is a transparent housing so that the surgeon canvisualize when tissue has deposited on the filter.

A suction trigger 816 couples to a suction port 818 for controllingsuction, with the port 818 being a source of suction for device 800.When the suction trigger is pulled back, the suction port opens. Whenthe suction port is connected to a vacuum source 819 the suction iscoupled through the filter and hollow shaft to the biopsy jaws. Thistransports any loosened tissue pieces from the biopsy jaws back to andtrapping them in the filter. Any fluid that is suctioned into the hollowshaft will pass through filter 820 into the filter housing and outthrough the suction port. Once the sample has been deposited on thefilter, the filter housing is rotated up and opened. The surgeon canthen remove valve cap 822 and the filter (the filter assembly) from thefilter housing. This filter assembly is placed into a container fortransport to the pathology laboratory. Another filter assembly can beinserted into the filter housing to collect more samples. The valve caphas a one-way valve, such as duck bill valve 824, preferably made ofsilicone, which allows for one way passage of suction from the biopsyjaws onto the filter. Once the filter assembly is placed into thecontainer for transport, the contents of the container cannot leak outthrough the one way valve.

Sectionable Filter adapted for histological laboratory use and manualloading.

Additional uses for the sectionable filter are shown in FIGS. 39 and 40,whereby the filter is used in the pathology lab to separate tissuesamples from the fixative or body fluids which may come to the lab fromany number of sources. This adapted sectionable filter can be configuredidentically to the ones integral to the biopsy collection container andused in conjunction with a standard prior art tissue cassette frame, orcould be configured as a sectionable filter cassette in a stage cassetteframe (rectangular version). Currently, these small tissue samples infixative are separated using a "tea bag" filter which separates thefluid from small tissue fragments. The tea bag then goes into the tissuecassette and the processor. When removed from the processor, the tissuefragments have become dried and are usually adhered to the tea bag,which requires scraping them loose and further manipulation to get themplaced into the paraffin mold form.

The sectionable lab filter or cassette configuration as shown in FIGS.39 and 40 is be adapted for use with a suction device 902 that can drawthe fluid through the filter quickly leaving tissue fragments 903 infilter 904 for cell block processing. The effluent could also be trappedin a cytospin container 905 inside the vacuum chamber 906 to make acytospin cytologic preparation.

FIG. 40 shows a sectional view of laboratory device 901 described above.Funnel 907 is attached to the instrument stand 908 and is adapted forplacement of a sectionable filter 904 or the cassette configuration 904in the central bore. Suction container 906 below the filter is threadedat 909 for attachment to the stand, a vacuum fitting 910 is incommunication with the inside of vacuum container 906. In use, a biopsysample arrives in a transport container 911 in fixative solution. Thecap is removed from the container and solution 912 is dispensed intofunnel 907. Large samples 903 and small samples 913 are strained throughsectionable filter 904 or sectionable filter cassette 904. Vacuum 902may be applied at this stage to speed up the process. The solution withsmaller fragments 913 passes through the filter and can be collected ina cytospin container 905 below the sectionable filter. The sectionablefilter or sectionable filter cassette is removed from the lab device andprocessed in a manner described herein.

Sectioned Paraffin Block

In general, the finished product is a sample of a tissue for analysiscomprising: means for supporting histologic tissue biopsy samplesincluding a microtome sectionable tissue supporting means for supportingtissue samples during tissue processing, embedding and microtomyincluding means for permitting said tissue supporting means to besuccessfully sectioned in a microtome, means for resisting histologicalstains, means for resisting degredation from solvents and chemicals usedto process and stain the tissue, and means for maintaining the tissuesupporting means non-distracting during tissue preparation and slidepreparation; and a supporting surface for supporting the sample formicroscopic examination. One form of the invention includes a poroustissue supporting means.

A finished product is specifically illustrated in FIGS. 41-46. Thus, inFIG. 41, a finished cassette 1000 has a tissue 1002 and tissuesupporting means 1004 and 1005' embedded in wax 1005; while FIG. 42shows a cassette 1000' which has wax 1006 embedding a tissue 1008 heldon pegs or posts 1010 in a manner similar to that indicated in FIG. 3.

Slicing the wax embedded tissue and tissue supporting means (filter) isindicated in FIGS. 43 and 44. It is emphasized that the filter materialis sectioned in the microtome along with the wax and the tissue sample.Thus, in FIG. 43, a microtome 1012 has a cassette 1014 thereon with atissue specimen indicated at 1016. A further view is shown in FIG. 44with tissue being indicated at 1018, wax at 1020, filter material(tissue supporting material) at 1022 and the cassette being indicated at1024. The microtome blade is indicated at 1026 as it slices thewax/tissue/filter combination. A mounted specimen is shown in FIG. 45with the sliced filter being shown at 1030 and the sliced tissuespecimen being shown at 1032 and the mounting slide being shown at 1034.Wax is indicated at 1035. A tissue specimen 1036 is shown in FIG. 46with holding posts 1040 and wax 1042 on a supporting means, such asslide 1044, for supporting the sample for microscopic examination.

In summary, the key components of the present invention include thefollowing.

1. The invention of tissue trapping filters or stages including thosethat are microtome sectionable or not and those that act as filters aswell as those that act as immobilizing stages; all can have a verticallytranslatable sample surface within a cassette frame which facilitatessample loading, confers protection from crushing of the tissue samplesduring the processing steps and allows the sample surface to be pushedinto the wax mold; use of the tissue trapping platforms (filter or stagein combination with a cassette frame) allows the tissue processing andwax embedding procedures to be automated.

2. An immobilization process, whereby the tissue is secured to a filteror stage by various means (Dry Net, Ballistic Net, etc.) allowing it tobe properly oriented for sectioning at the initial gross in, whicheliminates the need for further handling of the samples during tissueprocessing and wax embedding and therefore makes automation of theseprocesses possible.

3. Proper orientation of tissue samples is assured throughout theprocess.

4. The invention of sample trapping containers which contain asectionable filter and help to preserve the quality of the sample fromcollection to gross in and again reduce the amount of handling requiredfor the samples.

5. The invention of a Fine Needle Aspiration Device and needleconfigurations which can be used with the sectionable filter;

6. The invention of a surgical biopsy device with integral tissuetrapping sectionable filter.

7. The automation of the gross in procedure.

8. The automation of the tissue processing and wax embedding processestogether.

9. The automation of the dispensing of Fine Needle Aspiration Biopsy aswell as mucosal scrapings, endometrial curettes, bristle brush scrapingsetc., with collection of larger tissue pieces onto a sectionable filterand if desired the collection of eluate into a cytospin container forcytology.

10. A method for conducting tests on histology tissue biopsy samplescomprising: removing a tissue sample from a patient; placing the tissuesample onto a support, which can be microtomable if desired and whichcan, in one form of the invention, be porous; immobilizing the tissuesample on the support; subjecting both the support and the tissue sampleimmobilized thereon to a process for replacing tissue fluid with wax andimpregnating the tissue sample with wax, embedding the tissue sample ina wax mold to form a solid block of wax, using a microtome, slicing thesolid block of wax into thin slices; and mounting at least one of thethin slices on a support member for examination. If the tissuesupporting means is microtomable, then this element, along with thetissue sample, can be embedded in the wax, and both the sample and thesupport can be sliced by the microtome when the microtome slices theblock of wax.

By way of example, the above-discussed methods of obtaining tissuesamples is repeated herein along with the preferred form of tissuesupporting means:

Fine Needle Aspiration Biopsy--very small pieces of tissue taken fromthe core of a fine needle; usually transported in fixative solution;decant off fixative solution through a sectionable filter (180 μmfilter);

GI biopsy--characterized by a few small tissue pieces; it is desirableto concentrate the tissue pieces in close proximity to eachother--decant off fixative solution through a sectionable filter (1/4 mmfilter);

Prostate chips--orientation is irrelevant for these samples--sectionablefilter (1mm filter);

Endometrial Curettings--characterized by varying size samples;orientation is irrelevant--sectionable immobilization stage (1/2 mmfilter);

Vessel--orientation is critical; sections need to betransverse--sectionable immobilizing stage--manually position oververtical pegs;

Core Biopsy--i.e. from the prostate--orientation is critical; the tissueshould lie flat all in the same plane--sectionable immobilization stage;

Gall bladder--orientation is critical--the tissue should be embedded onedge--sectionable immobilization stage;

Uterine Wall, breast or large tumors--orientation is notcritical--sample lies flat in a plane--non-sectionable stage.

It is understood that while certain forms of the present invention havebeen illustrated and described herein, it is not to be limited to thespecific forms or arrangements of parts described and shown.

We claim:
 1. A means for supporting histologic tissue biopsy samplescomprising:a microtome sectionable tissue supporting means forsupporting tissue samples during tissue processing, embedding andmicrotomy including (1) means for permitting said tissue supportingmeans to be successfully sectioned in a microtome,(2) means forresisting histological stains, (3) means for resisting degredation fromsolvents and chemicals used to fix, process and stain the tissue and (4)means for maintaining said tissue supporting means non-distractingduring tissue processing and slide preparation.
 2. The means defined inclaim 1 wherein said tissue supporting means further includes means forresisting chemicals used in replacing tissue fluids with wax.
 3. Themeans defined in claim 2 wherein said tissue supporting means furtherincludes means for resisting molten wax used to embed the tissue sample.4. The means defined in claim 3 wherein said tissue supporting meansincludes a low density thermoplastic material.
 5. The means defined inclaim 3 wherein said tissue supporting means is porous and has a poresize of about 1 mm.
 6. The means defined in claim 3 wherein said tissuesupporting means is porous and has a pore size of about 1/4 mm.
 7. Themeans defined in claim 1 further including a collar surrounding saidtissue supporting means.
 8. The means defined in claim 7 furtherincluding a frame and means for connecting said tissue supporting meansto said frame.
 9. The means defined in claim 8 wherein said supportfurther includes projections on said collar.
 10. The means defined inclaim 9 wherein said frame further includes a first groove definedtherein for accommodating said projections.
 11. The means defined inclaim 10 wherein said frame further includes a second groove spaced fromsaid first groove.
 12. The means defined in claim 7 wherein said collaris microtome sectionable.
 13. The means defined in claim 1 wherein saidtissue supporting means is porous.
 14. The means defined in claim 1wherein said tissue supporting means is adapted to support tissue duringgrossing in.
 15. The means defined in claim 1 wherein said tissuesupporting means includes a tissue-immobilizing medium thereon.
 16. Themeans defined in claim 1 further including means for improving thewettability of said tissue supporting means.
 17. The means defined inclaim 1 wherein the means for maintaining said tissue supporting meansnon-distracting includes means for making the supporting means readilyidentifiable.
 18. The means defined in claim 1 wherein the tissuesupporting means further includes means for trapping a tissue sample.19. The means defined in claim 1 wherein the means for supporting tissuesamples includes means for maintaining tissue samples in an orientationwhereby all supported tissue samples are located in a common sectioningsurface.
 20. The means defined in claim 1 wherein the means forsupporting tissue samples includes means for engaging an automaticpicking device.
 21. The means defined in claim 1 wherein the means forsupporting tissue samples includes machine readable code thereon. 22.The means defined in claim 1 wherein the tissue supporting means furtherincludes a cover.
 23. The means defined in claim 1 wherein the tissuesupporting means further includes means for immobilizing a tissuesample.
 24. A means for supporting histologic tissue biopsy samplesincludinga microtome sectionable tissue supporting means for supportingtissue samples during tissue processing, embedding and microtomyincluding(a) means for permitting said tissue supporting means to besuccessfully sectioned in a microtome, (b) means for resistinghistological stains, (c) means for resisting degredation from solventsand chemicals used to process and stain the tissue, and (d) means formaintaining said tissue supporting means non-distracting during tissuepreparation and slide preparation; and a supporting means for supportingthe sample for microscopic examination.
 25. The means defined in claim24 wherein said supporting surface is a microscope slide.
 26. The meansdefined in claim 24 wherein said tissue supporting means is adapted tosupport tissue during grossing in.
 27. The means defined in claim 24wherein said tissue supporting means includes a tissue-immobilizingmedium thereon.
 28. A means for supporting histologic biopsy samplescomprising:a microtome sectionable tissue supporting means forsupporting tissue samples during tissue processing, embedding andmicrotomy wherein said tissue supporting means is formed of a materialwhich can be microtomed without significantly dulling the microtomeblade and which resists histological stains and which resistsdegradation from solvents and chemicals used to fix, process and staintissue samples and which is non-distracting with respect to the tissuesamples during microscopic examination.
 29. The means defined in claim24 wherein the means for maintaining said tissue supporting meansnon-distracting includes means for making the tissue readilyidentifiable.
 30. The means defined in claim 24 wherein the tissuesupporting means further includes means for trapping a tissue sample.31. The means defined in claim 24 wherein the means for supportingtissue samples includes means for supporting tissue samples in anorientation whereby all supported tissue samples can be cut by themicrotome in a single pass.
 32. The means defined in claim 24 whereinthe means for supporting tissue samples includes means for engaging anautomatic picking device.
 33. The means defined in claim 24 wherein themeans for supporting tissue samples includes machine readable codethereon.
 34. The means defined in claim 24 wherein the tissue supportingmeans further includes a cover.
 35. The means defined in claim 24wherein the tissue supporting means further includes means forimmobilizing a tissue sample.